Use of pseudopterosins for promoting wound healing

ABSTRACT

Methods of promoting would healing and the growth and proliferation of keratinocytes, fibroblasts and endothelial cells are disclosed. These methods comprise contacting a wound with an effective wound healing amount of a composition comprising a pseudopterosin or pseudopterosin derivative.

RELATED APPLICATIONS

This application is a 371 of PCT/US96/09039, filed Jun. 6, 1996 which isa Continuation-in-Part of U.S. patent application Ser. No. 08/486,359,filed on Jun. 7, 1995, now U.S. Pat. No. 5,597,808 the entire teachingsof which are hereby incorporated into this application by reference.

BACKGROUND OF THE INVENTION

Complications are a constant risk with wounds that have not fully healedand remain open. Although most wounds heal quickly without treatment,some types of wounds resist healing. Wounds which cover large surfaceareas also remain open for extended periods of time. Consequently, itwould be advantageous to accelerate the wound healing process. However,present methods of promoting wound healing are inadequate.

Full and partial thickness burns are an example of a wound type whichoften covers large surface areas and therefore requires prolongedperiods of time to heal. As a result, life-threatening complicationssuch as infection and loss of bodily fluids often arise. In addition,healing in burns is often disorderly, resulting in scarring anddisfigurement. In some cases wound contraction due to excessive collagendeposition results in reduced mobility of muscles in the vicinity of thewound. Therefore, there is a continued need to accelerate the rate ofhealing of the burns and to promote healing processes that result inmore desirable cosmetic outcomes and less wound contraction andscarring.

Severe burns which cover large areas are often treated by skinautografts taken from undamaged areas of the patient's body. However,skin grafts suffer from low take rates and therefore often provide onlyshort-term coverage of burns. Consequently, there is also a need for newmethods which improve the acceptance rate of skin autografts.

Dermal ulcers are an example of wounds that resist healing.Consequently, dermal ulcers often become chronic wounds. For example,one in seven individuals with diabetes develop dermal ulcers on theirextremities, which are susceptible to infection. Individuals withinfected diabetic ulcers often require hospitalization, intensiveservices, expensive antibiotics, and, in some cases, amputation. Dermalulcers, such as those resulting from venous disease (venous stasisulcers), excessive pressure (decubitus ulcers) and arterial ulcers alsoresist healing. The current treatments are limited to keeping the woundprotected, free of infection and, in some cases, to restore blood flowby vascular surgery. Therefore, there is also a need to provide methodswhich accelerate the rate of the healing of chronic dermal skin ulcersbefore the onset of infection and without the need for expensive andinvasive treatments such as surgery.

SUMMARY OF THE INVENTION

The present invention is based on the discovery that pseudopterosins orpseudopterosin derivatives accelerate the healing of a wide variety ofwounds. It is now been found that when applied to a wound,pseudopterosins or pseudopterosin derivatives promote the growth andproliferation of keratinocytes, fibroblasts and endothelial cells. Theyalso cause collagen matrix formation in the wound, which can result inreduced scarring. Based on this discovery, methods for promoting thehealing of wounds are disclosed.

One embodiment of the present invention is a method of promoting thehealing of a wound in an individual or animal. The method comprisescontacting the wound with a composition comprising an effective woundhealing amount of a compound having the structure in Formula I: ##STR1##wherein:

R1 and R2 are selected from the group consisting of --H, lower alkyl,substituted lower alkyl, phenyl, substituted phenyl, benzyl, substitutedbenzyl, acyl, alkoxycarbonyl and a monosaccharide, and wherein R1 and R2must be different, and one of R1 or R2 is a monosaccharide; and

R3 is a substituted or unsubstituted lower alkyl group.

Another embodiment of the present invention is a method of promoting thegrowth and proliferation of keratinocytes, fibroblasts and endothelialcells in a wound on an individual or animal. The method comprisescontacting the wound with a composition comprising an effective woundhealing amount of a compound having the structure of Formula I.

The methods of the present invention are useful in accelerating the rateof wound healing on an individual or an animal. Shortened healing timescan reduce the risk of infection, can reduce the losses of body fluidsfrom burn wounds, and can result in fewer complications such ascellulitis, osteomyelitis and gangrene in diabetic patients. Acceleratedwound healing can also bring about wound closure in patients withchronic skin ulcers. Better clinical outcomes can be obtained when themethods of the present invention are used to promote wound healing, e.g.reduced scarring, disfigurement and skin contraction.

DESCRIPTION OF THE FIGURES

FIG. 1 is a graph illustrating the wound healing effect ofpseudopterosin A methyl ether in a cream formulation compared withcontrols on skin lesions which had been chemically induced on the backsof guinea pigs.

FIG. 2 is a graph illustrating the wound healing effect ofpseudopterosin A methyl ether in an ointment compared with controls onskin lesions which had been chemically induced on the backs of guineapigs.

DETAILED DESCRIPTION OF THE INVENTION

The method of present invention can be used to promote the healing of awide variety of wounds, including wounds to external epithelial tissue,internal epithelial tissue, dental tissue and eye tissue. Wounds toexternal epithelial tissue are preferred and are of several types:excisional, burns, dermal skin ulcers, lesions due to dermatologicaldiseases, and atopic dermititus due to immediate type hypersensitivity.

Excisional wounds include tears, cuts, punctures or lacerations in theepithelial layer of the skin and may extend into the dermal layer andeven into subcutaneous fat and beyond. Excisional wounds can result fromsurgical procedures or from accidental penetration of the skin.

Burn wounds refer to cases where large surface areas of skin have beenremoved or lost from an individual. The loss of skin refers to theepidermal layer, and usually includes the dermal layer. Wounds of thistype include cases where part of the dermis has been lost or where thewound penetrates to subcutaneous fat and beyond. A burn wound can alsoresult when an individual's skin is exposed to a chemical agent.Typically, burn wounds result when a large area of an individual's skinis exposed to heat, such as in a fire. As used herein, burn wounds alsoinclude cases where large areas of the skin have been removed throughabrasion or surgically, e.g. to remove a skin cancer or to provide askin autograft.

Dermal skin ulcers refer to lesions on the skin caused by superficialloss of tissue, usually with inflammation. Dermal skin ulcers which canbe treated by the method of the present invention include decubitusulcers, diabetic ulcers, venous stasis ulcers and arterial ulcers.Decubitus wounds refer to chronic ulcers that result from pressureapplied to areas of the skin for extended periods of time. Wounds ofthis type are often called bedsores or pressure sores. Venous stasisulcers result from the stagnation of blood or other fluids fromdefective veins. Arterial ulcers refer to necrotic skin in the areaaround arteries having poor blood flow.

"Dental tissue" refers to tissue in the mouth which is similar toepithelial tissue, for example gum tissue. Thus, the method of thepresent invention is useful for treating periodontal disease. "Internalepithelial tissue" refers to tissue inside the body which hascharacteristics similar to the epidermal layer in the skin. Examplesinclude the lining of the intestine. Consequently, the method of thepresent invention is useful for promoting the healing of certaininternal wounds, for example wounds resulting from surgery. A "wound toeye tissue" refers to severe dry eye syndrome, corneal ulcers andabrasions and ophthalmic surgical wounds.

Wounds caused by dermatological diseases include lesions resulting fromautoimmune disorders such as psoriasis. Atopic dermititis refers to skintrauma resulting from allergies associated with an immune responsecaused by allergens such as pollens, foods, dander, insect venoms andplant toxins.

A method which "promotes the healing of a wound" results in the woundhealing more quickly as a result of the treatment than a similar woundheals in the absence of the treatment. "Promotion of wound healing" canalso mean that the method causes the proliferation and growth ofkeratinocytes, fibroblasts and endothelial cells, or that the woundheals with less scarring, less wound contraction, less collagendeposition and more superficial surface area. In certain instances,"promotion of wound healing" can also mean that certain methods of woundhealing have improved success rates, (e.g. the take rates of skingrafts,) when used together with the method of the present invention.The method can promote the healing of wounds on humans and animals suchas dogs, farm animals, guinea pigs, cats and the like.

The composition used in the present invention to promote wound healingcomprises an effective wound healing amount of a pseudopterosin or apseudopterosin derivative. The pseudopterosins are a class of naturalproducts isolated from the sea whip Pseudopterogorgia (Look et al., J.Org. Chem. 51:5140 (1986). These compounds are structurally related toaglycon (1), having an aldose sugar bonded to one of the phenols of thetricyclic ring system of (1) through a glycoside linkage at is anomericcenter. A numbering system for the ring carbons of the aglycon isindicated in (1). ##STR2##

Examples of pseudopterosins include pseudopterosins A-D (2-5), whichwere the first compounds of this class to be isolated (Look et al., J.Org. Chem. 51:5140 (1986). Recently, a series of pseudopterosins havingstructural differences from pseudopterosins A-D were isolated from P.elisabethae (Roussiss et al., J. Org. Chem. 55:4916 (1990)). The sugarmoiety in pseudopterosin E (6) and F (7) is attached to the phenol atcarbon 10 of the aglycon instead of the phenol at carbon 9, as inpseudopterosins A-D. In pseudopterosins G-J (8-11), the stereochemicalconfiguration at carbon 7 of the aglycon is inverted with respect topseudopterosins A-D. The aglycon of pseudopterosins K (13) and L (14) isenantiomeric to the aglycon of pseudopterosins A-D. Pseudopterosinderivatives are also useful for the methods described herein. As usedherein, a pseudopterosin derivative is a pseudopterosin in which theaglycon (1) and/or the monosaccharide substructures are chemicallymodified and which promotes wound healing. Examples of pseudopterosinderivatives include pseudopterosins in which-the free phenol of theaglycon is alkylated or acylated and are referred to herein as"pseudopterosin alkyl ethers" or "acylated pseudopterosins",respectively. Specific examples of pseudopterosin A derivatives includepseudopterosin A methyl ether (15) (R2=--CH₃), pseudopterosin A4-hydroxybutyl ether (R2=(--CH₂)₄ --OH)), pseudopterosin A pentyl ether(R2=--(CH₂)₄ --CH₃), pseudopterosin A acetamide ether (R2=--CH₂CO--NH₂), and pseudopterosin A benzyl ether (R2=--CH₂ --C₆ H₅).Pseudopterosin derivatives also include pseudopterosins wherein thehydrocarbon side chain attached to carbon one is modified, for exampleby hydrogenation, or by oxidation of the 2-methyl-1-propene moiety to,for example, 1-keto-2-methyl-propane or 2-methyl-propeneoxide (Jacobs,et al., U.S. Pat. No. 4,849,410). Chain elongation of these ##STR3##oxidized pseudopterosins can be carried out by methods known to thoseskilled in the art.

In one embodiment the pseudopterosin or pseudopterosin derivative hasthe structure of Formula II: ##STR4##

R1 and R2 are each selected from the group consisting of --H, loweralkyl, substituted lower alkyl, phenyl, substituted phenyl, benzyl,substituted benzyl, acyl, alkoxycarbonyl and a monosaccharide. R1 and R2must be different and one of R1 and R2 is the monosaccharide.

R3 is a substituted or unsubstituted lower alkyl group. Preferably, R3is --CH═C(CH₃)₂ or --CH₂ CH(CH₃)₂.

As used herein, "lower alkyl" refers to a hydrocarbon containing one toabout ten carbon atoms. The hydrocarbon can be saturated or can containone or more units of unsaturation. The hydrocarbon can also be branchedor straight chained.

Suitable substituents on a lower alkyl group include hydroxy, alkoxy,ketone, aldehyde, amide (--CO--NH₂), alkyl ester, alkyl amine, benzyl,substituted benzyl, phenyl, substituted phenyl, amine and the like.Suitable substituents on a phenyl or benzyl group include nitro, cyano,C1-C4 straight chain or branched alkyl, halo, alkoxy and the like.

An "acyl" group is --C(O)-(lower alkyl), wherein alkyl is as definedabove. An "alkoxy carbonyl" group is --C(O)--O-(lower alkyl), whereinlower alkyl is as defined above.

The aglycon portion of the compound of the present invention has fourchiral centers, resulting in sixteen possible stereoisomers. Preferredstereoisomers are those in which the stereochemical configurations atthe four chiral carbons in the aglycon are the same as in pseudopterosinA (1), pseudopterosin G (8) or pseudopterosin K (13). The stereochemicalconfiguration of pseudopterosin A is most preferred.

As used herein, a "monosaccharide" is a chiral polyhydroxy aldehyde in acyclic hemiacetal form (referred to as an "aldose") or a chiralpolyhydroxy ketone in a cyclic hemiketal form (referred to as a"ketose"). Chiral polyhydroxy aldehydes are preferred. Examples ofsuitable polyhydroxy aldehydes include aldotrioses, aldotetroses (e.g.D- and L-erythrose and threose), aldopentoses (e.g. D- and L-arabinose,xylose, ribose, and lyxose), aldohexoses (e.g. D- and L-glucose, allose,altrose, mannose, gulose, idose, galactose and talose), aldoheptoses,aldo-octoses and aldononoses. The polyhydroxy aldehyde can be in afuranose form (for aldoses having four carbons or more) or pyranose form(for aldoses having five carbon atoms or more). The pyranose form ispreferred.

Suitable polyhydroxy ketones include pentuloses, hexuloses (e.g.D-fructose, D-sorbose, D-psicose and D-tagatose), heptuloses, octulosesand nonuloses. The polyhydroxy ketone can be in a furanose form (forketoses having five carbons or more) or pyranose form (for ketoseshaving six or more carbon atoms). The pyranose form is preferred.

A "monosaccharide" can also include the monosaccharides describedhereinabove in which one or more of the hydroxy groups arefunctionalized with an alcohol protecting group. More than one kind ofalcohol protecting group can be used in a single monosaccharide.Suitable alcohol protecting groups include lower alkyl (preferablymethyl), acyl (preferably acetyl), benzoyl, substituted benzoyl(suitable substituents include, for example, lower alkyl, nitro, halide,alkoxy and cyano) and carbonates. Other suitable protecting can be foundin Greene and Wuts, "Protective Groups in Organic Synthesis," secondedition, John Wiley-and Sons, Inc., 1991. One or more hydroxy groups onthese monosaccharides can be protected with alcohol protecting groups,as described above. The preferred alcohol protecting group is acetyl.

In a preferred embodiment the monosaccharide has the structure ofStructural Formula III: ##STR5##

R4, R5 and R6 are independently selected from the group consisting of--H and an alcohol protecting group. Suitable alcohol protecting groupsare described above. A preferred alcohol protecting group is acyl, andis even more preferably acetyl.

R7 is selected from the group consisting of --H, --CH₃ and --CH₂ OR8. R8is selected from the group consisting of --H, a lower alkyl group and analcohol protecting group. Suitable lower alkyl and alcohol protectinggroups are as described above. Acyl is a preferred alcohol protectinggroup, and acetyl is even more preferred.

In one aspect R1 is the monosaccharide, R3 is --CH═C(CH₃)₂ or --CH₂--CH(CH₃)₂, R4, R5 and R6 are each independently --H or acyl (preferablyacetyl), and R7 is selected from the group consisting of --H, --CH₃ and--CH₂ OH. Examples include compounds wherein the aglycon has thestructure of pseudopterosin A (1), pseudopterosin G (8) andpseudopterosin K (13) and R1 is α-L-fucose, α-D-arabinose or β-D-xylose.One or more of the hydroxy groups on the monosaccharide moiety can beprotected with an acyl group, preferably acetyl. Alternatively, R2 isthe monosaccharide. Examples include compounds wherein the aglycon hasthe structure of pseudopterosin E (6) and the R2 is α-L-fucose,α-D-arabinase or β-D-xylose.

In a more preferred embodiment of the present invention, the compoundwhich promotes wound healing has the structure of Structural Formula IV:##STR6##

R1 has the structure of Structural Formula V: ##STR7##

R2 is selected from the group consisting of --H, lower alkyl, benzyl,--(CH₂)_(m) -benzyl, --(CH₂)_(m) -(substituted benzyl), --(CH₂)_(m)--CH₂ OH and --(CH₂)_(m) --CONH₂, wherein m is an integer from 1 toabout 9; m is preferably an integer from 1 to 4. Preferred lower alkylgroups are C1-C6 straight chain, saturated hydrocarbons. It is mostpreferred that R2 is methyl or --H. R3 is --CH═C(CH₃)₂. R4, R5 and R6are each independently selected from the group consisting of --H andacetyl. R7 is independently selected from the group consisting of --H--CH₃ and --CH₂ OH. It is most preferred that R4, R6 and R7 are each--H, i.e. the compound is pseudopterosin A, pseudopterosin A methylether, pseudoptersoin C or pseudopterosin C methyl ether.

In another more preferred embodiment the compound which promotes woundhealing has the structure of Structural Formula IV. R1 is selected fromthe group consisting of --H, lower alkyl, benzyl, --(CH₂)_(m) -benzyl,--(CH₂)_(m) -substituted benzyl, --(CH₂)_(m) --CH₂ OH and --(CH₂)_(m)--CONH₂, wherein m is an integer from 1 to about 9. m is preferably 1-4.Preferred lower alkyl groups are C1-C6 straight chain saturatedhydrocarbons. It is most preferred that R1 is methyl or --H. R2 has thestructure of Structural Formula VI: ##STR8## R3 is --CH═C(CH₃)₂. R4, R5and R6 are each independently selected from the group consisting of --Hand acetyl. --H is most preferred. R7 is independently selected from thegroup consisting of --H --CH₃ and --CH₂ OH. --CH₃ is most preferred.

Other examples of suitable compounds include pseudopterosin A benzylether, pseudopterosin A o-nitrobenzyl ether, pseudopterosin A pentylether, pseudopterosin A decyl ether, pseudopterosin A octadecyl ether,pseudopterosin A 4-hydroxy butylether, pseudopterosin A acetamide(wherein R2 in Formula IV is --CH₂ --NH₂), pseudopterosin Atetraacetate, pseudopterosin B, pseudopterosin D, pseudopterosin Bmethyl ether, pseudopterosin D methyl ether, pseudopterosin A ethylether, pseudopterosin B ethyl ether, pseudopterosin C ethyl ether,pseudopterosin D ethyl ether, acetyl pseudopterosin A, acetylpseudopterosin B, acetyl pseudopterosin C, acetyl pseudopterosin D,pseudopterosin E ethyl ether, acetyl pseudopterosin E, pseudopterosin F,pseudopterosin F methyl ether, pseudopterosin F ethyl ether, acetylpseudopterosin F, pseudopterosin G, pseudopterosin G methyl ether,pseudopterosin G ethyl ether, acetyl pseudopterosin G, pseudopterosin H,pseudopterosin H methyl ether, pseudopterosin H ethyl ether, acetylpseudopterosin H, pseudopterosin I, pseudopterosin I methyl ether,pseudopterosin I ethyl ether, acetyl pseudopterosin I, pseudopterosin J,pseudopterosin J methyl ether, pseudopterosin J ethyl ether, acetylpseudopterosin J, pseudopterosin K, pseudopterosin K methyl ether,pseudopterosin K ethyl ether, acetyl pseudopterosin K, pseudopterosin L,pseudopterosin L methyl ether, pseudopterosin L ethyl ether and acetylpseudopterosin L.

Pseudopterosin A, C and E are isolated from Pseudopterogorgiaelisabethae according to known procedures (Look et al., J. Org. Chem.,51:5140 (1986) and Roussis et al., J. Org. Chem., 55:4916 (1990)). Alkylethers of pseudopterosins are prepared by alkylating the pseudopterosinwith an alkylating agent such as methyl iodide (Look et al., Roussis etal. and Jacobs et al. U.S. Pat. No. 4,849,410). Pseudopterosin B-L andalkylated derivatives thereof can be isolated and prepared by the samebasic procedures (Look et al. and Jacobs et al. U.S. Pat. No. 4,745,104,Jacobs et al. U.S. Pat. No. 4,849,410 and Roussis et al.). Methods ofpreparing pseudopterosin derivatives having other monosaccharides arecarried out by methods known in the art (Corey and Carpino, J. Am. Chem.Soc., 111:5472 (1989)). Examples of the preparation of otherpseudopterosin derivatives are given in Examples 4-9 and discussedfurther in U.S. patent application filed Jun. 7, 1995, now U.S. Pat. No.5,624,911 naming William H. Fenical and Robert S. Jacobs as inventors,the teachings of which are incorporated herein by reference.

The composition used in the present invention to promote wound healingcan additionally comprise an inert, non-toxic solvent such as acetone oralcohol in which the pseudopterosin or pseudopterosin derivative isdissolved, or, preferably, a pharmaceutical carrier suitable for localtopical administration in which the pseudopterosin or pseudopterosinderivative is dissolved or suspended. Examples of pharmaceuticallyacceptable carriers include, for example, commercially available inertgels, or liquids supplemented with albumin, methyl cellulose or acollagen matrix. Typical of such formulations are ointments, creams andgels. Ointments are typically prepared using an oleaginous base, e.g.,containing fixed oils or hydrocarbons, such as white petrolatum ormineral oil, or an absorbent base, e.g., consisting of an absorbentanhydrous substance or substances, for example anhydrous lanolin.Following formation of the base, the active ingredients are added in thedesired concentration. Creams generally comprise an oil phase (internalphase) containing typically fixed oils, hydrocarbons, and the like, suchas waxes, petrolatum, mineral oil, and the like, and an aqueous phase(continuous phase), comprising water and any water-soluble substances,such as added salts. The two phases are stabilized by use of anemulsifying agent, for example, a surface active agent, such as sodiumlauryl sulfate; hydrophilic colloids, such as acacia colloidal clays,beegum, and the like. Upon formation of the emulsion, the activeingredients are added in the desired concentration. Gels are comprisedof a base selected from an oleaginous base, water, or anemulsion-suspension base, as previously described. To the base is addeda gelling agent which forms a matrix in the base, increasing itsviscosity to a semisolid consistency. Examples of gelling agents arehydroxypropyl cellulose, acrylic acid polymers, and the like. The activeingredients are added to the formulation at the desired concentration ata point preceding addition of the gelling agent.

Preferred formulations are those which promote penetration of dermal andepidermal layers of the skin by the pseudopterosin or pseudopterosinderivative and in which the pseudopterosin or pseudopterosin derivativeis stable. A preferred formulation is a petrolatum ointment comprisingfrom between about 0.005% to about 2.0% of the pseudopterosin orpseudopterosin derivative, e.g. pseudopterosin A methyl ether. Theointment is prepared according to the procedure in Example 3 and has thefollowing composition:

    ______________________________________                                                       % W/W                                                          ______________________________________                                        White Petrolatum, USP                                                                          82.5-84.5                                                    White Wax, NF    10.0                                                         Cholesterol, NF  3.0                                                          Diisopropyl Adipate                                                                            2.5                                                          Pseudopterosin   0.005-2.0                                                    ______________________________________                                    

Another preferred formulation is an oil/water cream with between about0.005% and about 2.0% pseudopterosin or pseudopterosin derivative havingthe following composition:

    ______________________________________                                        Stearic Acid            8.0%                                                  Cetyl Alcohol           0.8%                                                  Polyoxyethylene Laurel Ether (BRIDGE 30 ™)                                                         1.5%                                                  Octyldodecanol          14.4%                                                 Carboxyvinyl Polymer (CARBOMER 980 ™)                                                              0.6%                                                  Sodium Laureth Sulfate  0.3%                                                  Sodium Hydroxide        1.2%                                                  Methylparaben           0.2%                                                  Propylparaben            0.05%                                                Pseudopterosin          0.005-2.0%                                            Purified Water          70.95-72.95%                                          ______________________________________                                    

Both of these preferred formulations with 0.5% pseudopterosin A methylether provide stability greater than six months at room temperature.

The wound being treated is contacted with a composition comprising aneffective wound healing amount of a pseudopterosin or pseudopterosinderivative, for example by applying the composition directly to thewound. In the case of wounds to eye tissue, the compound can optionallybe applied by eye drops.

An "effective wound healing amount of a pseudopterosin or apseudopterosin derivative" contains a sufficient quantity of thepseudopterosin or a pseudopterosin derivative to promote wound healingand the growth and proliferation of endothelial cells, keratinocytes andfibroblasts. The skilled artisan will appreciate that the quantity ofpseudopterosin or a pseudopterosin derivative which promotes woundhealing depends on the specific nature of the wound, e.g. wound type andseverity, and can vary the amount of compound used, depending on theapplication. The amount of pseudopterosin or pseudopterosin derivativeapplied to the wound depends on the amount of the composition (e.g.,inert solvent and pharmaceutical carrier) applied to the wound and theconcentration of the pseudopterosin or pseudopterosin derivative in thepharmaceutical carrier or inert solvent. Generally, enoughpharmaceutical carrier or inert solvent is used to cover the wound.Suitable concentrations of pseudopterosin or pseudopterosin derivativein the pharmaceutical carrier or inert solvent ranges from about 0.005%to about 2.0%, preferably from about 0.05% to about 0.5%.

In certain instances where wounds are being treated, it may beadvantageous to co-administer one or more additional pharmacologicallyactive agents to the wound along with the compounds of the present ofinvention. For example, infection is a threat with any open wound,particularly in burn wounds. One aspect of the present invention is toco-administer to the wound an antimicrobial, a disinfectant or anantibiotic. Managing pain and inflammation are also important aspects oftreating wounds. The compounds of the present invention can also beco-administered to the wound along with a pain-relieving agent such asan analgesic or an anti-inflammatory agent.

Another embodiment of the present invention is a method of improving thetake rate of a skin graft. Because there is no dermis or basementmembrane present at the time an epithelial autograft is applied to awound, there is nothing to secure the autograft to the wound bed.Consequently, autografts tend to blister and shear, decreasing thelikelihood that the autograft will "take", i.e. adhere to the wound andform a basement membrane with the underlying granulation tissue. Takerates can be increased by providing a physiological scaffolding in thewound onto which keratinocytes and endothelium cells can migrate and bypromoting the formation and migration of capillaries into thescaffolding. Pseudopterosin A methyl ether has been observed (Example 1)to promote the growth and proliferation of fibroblasts and theneovascularization of wounds. Increased collagen biosynthesis anddeposition results, thereby forming a collagen matrix scaffolding withinthe wound. Consequently, the application of a pseudopterosin orpseudopterosin derivative to a wound also promotes the processesnecessary to increase the take rates of skin autografts. The method ofincreasing take rates comprises contacting the skin autograft with aneffective wound healing amount of the compounds and compositionsdescribed in the method of promoting wound healing and in the method ofpromoting the growth and proliferation of keratinocytes, fibroblasts andendothelial cells in a wound, as described above.

The invention is further illustrated by the following examples, whichare not intended to be limiting in any way.

EXEMPLIFICATION EXAMPLE 1

Healing of Full Thickness Lesions on Hartley Guinea Pigs Promoted byPseudorterosin A Methyl Ether

Full thickness lesions were induced on the backs of Hartley Strainguinea pigs (Skoog, M. L. Acta Derm Venerol (Stockh) 60(3):239-44, 1980)by the cutaneous application of 1-chloro-2,4-dinitrobenzene (DNCB).Shaved Hartley male guinea pigs were treated once daily for four dayswith 1-2% DNCB in acetone to induce the full thickness wounds involvingboth dermal and epidermal portions. The DNCB caused the loss of theepidermis thus subjecting the dermis to conditions not suitable for themaintenance of continued viability. Dermal appendages were necrotic,degenerating or lost. The remnants of the dermis were found to beacellular and edematous. These severe defects encompassed a significantarea of the demarcated region of irritant application and lackedintegrity of the epithelial barrier. The lesions and treatment area wereapproximately 9 cm². Animals in each group were treated with a creamformulation (See Example 2) containing 0.005%, 0.01%, 0.05%, 0.1% or0.5% pseudopterosin A methyl ether. Control,groups in which the animalswere treated with 0.05% fluocinonide (commonly sold under the trade nameLIDEX™), a cream placebo or were left untreated were also included.

In a second set of experiments, animals were treated with an ointment(See Example 3) containing 0.1% or 0.05% methyl pseudoterosin A. Animalstreated with an ointment placebo or left untreated were used ascontrols.

On days 5-9 animals were scored on a blind basis by two independentobservers using the Draize Criteria (Draize, J. H., Dermal Toxicity,"Appraisal of the Safety of Chemicals in Food, Drugs, and Cosmetics,"The Association of Food and Drug Officials of the United States, 46-59,1959). The Draize Criteria reflects erythema, edema, and other clinicalparameters. Control and treatment groups contained six animals. The meanvalue reflects the Draize score obtained on day 9. The results are shownin FIG. 1. Results were compared by unpaired, nonparametric analysis.

The effects of pseudopterosin A methyl ether (cream formulation) inreducing erythema and edema and in promoting regrowth of tissue werenoticeably superior to the effect of the steroid placebo controls. FIG.1 and FIG. 2 show that pseudopterosin A methyl ether in the creamformulation and in the ointment demonstrated a dose responserelationship with respect to its ability to decrease the DNCB-inducedinflammation. Placebo ointment and cream vehicles were found to lackclinical efficacy.

Skin harvested from the central portion of the lesions from non-treatedanimals or placebo controls revealed delamination and loss of theepidermis, necrosis of hair follicles, sweat glands and other dermalappendages, fibroblast necrosis, edema of the dermis and othermanifestations of severe full thickness skin lesions. The skin appearednon-viable since viable cellular elements were not observed. Clinicallyand histopathologically, the necrotic lesion induced by the chemicalirritant, coupled with the host response, resulted in tissue destructionand loss of the epidermis.

Microscopic examination of skin taken from animals treated withclobetasol propionate (Temovate™) was similar to untreated or placebotreated animals. Loss of the epidermis was confirmed histologically innearly all cases. The apical regions of the dermis were judged to benon-viable with remnants of inflammatory foci and necrotic fibroblasts.Dermal appendages including hair follicles, sebaceous and sweat glands,and other dermal components were noted to be necrotic or lost. Deeperzones of the dermis appeared to contain viable fibroblasts with minimal,patent vasculature. Considerable edema was noted in the dermis.

In contrast to the untreated or placebo treated animals, all animaltreated with pseudopterosin A methyl ether displayed the presence of ahypertrophic epidermis. Histopathological assessment of the hypertrophicepidermis was confirmed by the increase in the thickness of theepidermis, increased thickness of the stratum granulosum and thepresence of nuclear remnants within the stratum corneum. In addition,frequent mitotic figures were noted within the basal keratinocytes. Thedermis from animals treated with pseudopterosin A methyl ether displayedactive patterns of fibroblast proliferation, neovascularization, and newcollagen matrix deposition. This aggressive repopulation of the dermistook place within the context of existing dermal structure thus limitingor preventing apparent scar formation. The striking picture ofneovascularization, as evidenced by dilated capillary vasculature withprominent endothelium, indicates that an active process was mediated bypseudopterosin A methyl ether in these animals. Clinical observationconfirmed that epidermal regrowth including hair was accomplished byproliferation of dermal appendageal epithelial derivatives. In contrastto untreated animals having focal necrosis with active bleeding andconsiderable eschar, animals treated with pseudopterosin A methyl etherdisplayed nearly complete resolution of lesions.

Animals treated with pseudopterosin A methyl ether displayed significantand consistent evidence of wound healing. The final outcome clearlyreveals a continuous epithelial barrier and dermal component of skinthat is consistent with active cellular proliferative responses typicalof wound healing. Keratinocytes, dermal fibroblasts and capillaryendothelium are all recognized in patterns consistent with an activeresolution phase secondary to injury and tissue destruction. Thesecellular elements are proliferative and nuclear morphology indicatedconsiderable biosynthetic activity. Epidermis was noted to behypertrophic, another common feature of wound healing.

Increased matrix formation through collagen deposition typically resultsin reduced scarring (Ehrlich, J. of Trauma, 24(5):35 (1984) and Ehrlich,Prog. Clin. Biol. Res., 266:243 (1988)), suggesting that treatment ofwounds with a pseudopterosin or a pseudopterosin derivative gives abetter clinical outcome as well as accelerated healing rates. This wasconfirmed by visual observation of healed wounds which had been treatedwith pseudopterosin A methyl ether. As expected, these wounds werecharacterized by larger surface areas and decreased scarring.

EXAMPLE 2

Preparation of a Pseudopterosin Ointment Formulation

    ______________________________________                                                        % W/W                                                         ______________________________________                                        White Petrolatum, USP                                                                           84.0                                                        White Wax, NF     10.0                                                        Cholesterol, NF   3.0                                                         Diisopropyl Adipate                                                                             2.5                                                         Pseudopterosin    0.5                                                         ______________________________________                                    

Pseudopterosin A methyl ether was added to diisopropyladipate and mixedwell until dissolved. The mixture was sonicated to increase the speed ofdissolution. All other components were combined in a suitable vessel andheated to approximately 75° C. After all components are melted, thecombination was mixed with a propeller mixer while allowing to cool.While continuing to propeller mix, the pseudopterosin A methyl ethersubstance solution was incorporated into the ointment base when theointment had cooled to about 50° C. The mixture was cooled to roomtemperature, using intermittent hand stirring to ensure productuniformity.

EXAMPLE 3

Preparation of a 0.1% Ointment of Pseudopterosin A Methyl Ether

White petrolatum (82.4 grams), white wax (10.0 grams) and cholesterol(3.0 grams) were weighed, added to a stainless steel vessel and mixedwith a homogenizing mixer for five minutes at high speed. Mixing wascontinued at low speed while cooling to 50° C. by exposure to ambientair.

Diisopropyl adipate (2.5 grams) was weighed into a separate container.Pseudopterosin A methyl ether (0.1 grams) was added and mixed at 45° C.to 50° C. with stirring until completely dissolved. The warm diisopropyladipate solution was added to the ointment base at about 50° C. Theresulting batch was further mixed for five minutes with the homogenizingmixer at high speed. The batch was then allowed to cool below 30° C. ina room temperature water bath while mixing to maintain productconsistency.

EXAMPLE 4

Synthesis of Pseudopterosin A Benzyl Ether ##STR9## Preparation

Pseudopterosin A (150 mg in 50 ml acetone) was placed in a 100 ml flask.K₂ CO₃ (150 mg), sodium iodide (500 mg) and benzylchlorine (1.1 g=1 ml)were added. The solution was stirred and refluxed for 5 hours. Afterstirring overnight, 0.5 ml benzylchlorine was added and the solution wasrefluxed for another 6 hours. After cooling to room temperature, theyellow solution was concentrated by rotary evaporation. Water (50 ml)and CH₂ Cl₂ (30 ml) were added to the concentrate and the solution wastransferred to a separatory funnel and extracted 3 times with CH₂ Cl₂(3×50 ml). The total collected CH₂ Cl₂ layer were washed with brine(3×50 ml). Next, the solution was transferred to an Erlenmeyer flask anddried with Na₂ SO₄. The dry solution was filtered and concentrated undervacuum to give a yellow oil.

Purification

For purification, the yellow oil was chromatographed on a silica columneluting with 65/35 ethyl acetate/isooctane. The product was dried underhigh vacuum to give white crystals. Yield: 90 mg=50%.

EXAMPLE 5

Synthesis of Pseudopterosin A Pentyl Ether ##STR10## Preparation

Pseudopterosin A (150 mg in 70 ml acetone) was placed in a 100 ml flask.K₂ CO₃ (150 mg) and 1-iodopentane (1400 mg=0.92 ml) were added. Thesolution was stirred and refluxed overnight. After cooling to roomtemperature, the solution was concentrated by rotary evaporation. Water(30 ml) and CH₂ Cl₂ (30 ml) were added to the concentrate and thesolution was transferred to a separatory funnel and extracted 3 timeswith CH₂ Cl₂ (3×30 ml). The total collected CH₂ Cl₂ layers were washedwith brine (3×50 ml). Next, the solution was transferred to anErlenmeyer flask and dried with Na₂ SO₄. The dry solution was filteredand concentrated under vacuum to give a yellow oil.

Purification

For purification, the yellow oil was chromatographed on a silica columneluting with 50/50 ethyl acetate/isooctane. The product was dried underhigh vacuum to give white crystals. Yield: 84 mg=49%.

EXAMPLE 6

Synthesis of Pseudopterosin A Decanyl Ether ##STR11##

Pseudopterosin A (150 mg in 100 ml acetone) was placed in a 250 mlflask. K₂ CO₃ (150 mg) and 1-iododecane (930 mg -0.74 ml) were added.The solution was stirred and refluxed for 5 hours. After stirringovernight, 0.74 ml 1-iododecane and 150 mg K₂ CO₃ were added again andthe solution was refluxed for another 15 hours. After cooling to roomtemperature, the solution was concentrated by rotary evaporation. Water(50 ml) and CH₂ Cl₂ (50 ml) were added to the concentrate and thesolution was transferred in a separatory funnel and extracted 3 timeswith CH₂ Cl₂ (3×75 ml). The total collected CH₂ Cl₂ layers were washedwith brine (3×50 ml). Next, the solution was transferred to anErlenmeyer flask and dried with Na₂ SO₄. The dry solution was filteredand concentrated under vacuum to give a yellow oil.

Purification

For purification, the yellow oil was chromatographed on a silica columneluting with 50/50 ethyl acetate/isooctane. The product was dried underhigh vacuum to give white crystals. Yield: 105 mg=52%.

EXAMPLE 7

Synthesis of Pseudopterosin A Octadecanyl Ether ##STR12##

Pseudopterosin A (150 mg in 70 ml acetone) was placed in a 100 ml flask.K₂ CO₃ (1150 mg), sodium iodide (580 mg) and 1-bromooctadecane (2300 mg)were added. The solution was stirred and refluxed overnight. Afteradding 25 ml toluene, the solution was again placed in a 100 ml flask.K₂ CO₃ (1150 mg), sodium iodide (580 mg) and 1-bromooctadecane (2300 mg)were added. The solution was stirred and refluxed overnight. Afteradding 25 ml toluene, the solution was again stirred and refluxedovernight. Next, 25 ml water and 1000 mg of K₂ CO₃ were added, and thesolution was stirred and refluxed for another 5 hours. After cooling toroom temperature, the solution was concentrated by rotary evaporation.Water (50 ml) and CH₂ Cl₂ (30 ml) were added to the concentrate and thesolution was transferred in a separatory funnel and extracted 3 timeswith CH₂ Cl₂ (3×50 ml). The total collected CH₂ Cl₂ layers were washedwith brine (3×50 ml). Next, the solution was transferred into anErlenmeyer flask and dried with Na₂ SO₄. The dry solution was filteredand concentrated by rotary evaporation to give a yellow oil.

Purification

For purification, the yellow oil was chromatographed on a silica columneluting with 40/60 ethyl acetate/isooctane. The product was dried underhigh vacuum to give white crystals Yield: 100 mg=42%.

EXAMPLE 8

Synthesis of Pseudopterosin A Butanol Ether ##STR13## Preparation

Pseudopterosin A (150 mg in 50 ml acetone) was placed in a 100 ml flask.K₂ CO₃ (150 mg) and 4-iodobutyl acetate (1300 mg=0.8 ml) were added. Thesolution was stirred and refluxed overnight. Then, iodobutyl acetate(650 mg=0.4 ml) and water (2.5 ml) were added and the solution wasstirred and refluxed for another 5 hours. Then, 600 mg of KOH wereadded, and the solution was concentrated by rotary evaporation. Water(50 ml) and CH₂ Cl₂ (30 ml) were added to the concentrate and thesolution was transferred in a separatory funnel and extracted 3 timeswith CH₂ Cl₂ (3×50 ml). The total collected CH₂ Cl₂ layers were washedwith brine (3×50 ml). Next, the solution was transferred to anErlenmeyer flask and dried with Na₂ SO₄. The dry solution was filteredand concentrated under vacuum to give yellow oil.

Purification

For purification, the yellow oil was chromatographed on a silica columneluting with 65/35 ethyl acetate/isooctane. The product was dried underhigh vacuum to give white crystals. The product was not the expected PsAacetoxy butyl ether but the PsA butanol ether, derived by saponificationof the acetate under the basic conditions employed in the reaction.Yield: 46 mg=26%.

EXAMPLE 9

Synthesis of Pseudopterosin A Acetamide Ether ##STR14## Preparation

Pseudopterosin A (150 mg in 50 ml acetone) was placed in a 100 ml flask.K₂ CO₃ (1500 mg) and iodoacetamide (960 mg) were added. The solution wasstirred and refluxed for 5 hours. After cooling to room temperature, theyellow solution was concentrated by rotary evaporation. Water (50 ml)and CH₂ Cl₂ (30 ml) were then added to the concentrate and the solutionwas transferred in a separatory funnel and extracted 3 times with CH₂Cl₂ (3×50 ml). The total collected CH₂ Cl₂ layers were washed with brine(3×50 ml). Next, the solution was transferred into a Erlenmeyer flaskand dried with Na₂ SO₄. The dry solution was filtered and concentratedunder vacuum to give a yellow oil.

Purification

For purification, the yellow oil was chromatographed on a silica columneluting with 97/3 ethyl acetate/isooctane. The product was dried underhigh vacuum to give white crystals. Yield: 67 mg=40%.

EXAMPLE 10

Effects of Pseudopterosin A Methyl Ether Ointment on Normal PorcineModel

One 20 kg domestic Yorkshire Pig was used for the study described below.The animal received general inhalation anesthesia and was placed in aprone position. The dorsum was shaved and cleansed.

A series of 64×1 cm² grids was tattooed onto the dorsum of the animal.The dorsum was covered with an occlusive dressing and the animal wasawakened without difficulty.

After a 48 hour recovery period (to allow resolution of any skinirritation from creation of the grids), the pseudopterosin A methylether ointment was applied topically in concentrations of 0.05%, 0.15%and 0.5%. Vehicle alone and untreated skin served as controls. Ointmentwas applied to a 1 cm² square such that the surface area was completelycovered. Ointment was applied at daily intervals for five days.

Elliptical biopsies were taken at day 1, 3, 5 and 7 days postapplication of the ointment. Biopsies were bisected along the long axis.Tissue was placed in formaldehyde for histology (H&E) and frozen in OCTfreezing compound for further immunohistochemical analysis (ki-67staining for proliferation). An occlusive dressing was reapplied eachday after application of the ointment.

Gross observations of all treated and control sites revealednormal-appearing skin at all time points. No scaling, or erythema wasnoted. Histologic analysis revealed normal skin structure at all timepoints for all site biopsied. There were no signs of inflammatoryinfiltration or changes in epidermal architecture. Staining for Ki-67also revealed similar proliferative patterns across all treatmentgroups.

Topical application of pseudopterosin A methyl ether (at concentrationsof 0.05-0.5%) had no observable effect on normal porcine skin. Noirritation was observed, nor was any inflammation induced by applicationof pseudopterosin A methyl ether.

EXAMPLE 11

Ability of Pseudopterosin A Methyl Ether Ointment to PromoteRe-Epithelization of Partial Thickness Wounds

One 20 kg Yorkshire pig was used for the study described below. Theanimal received general inhalation anesthesia and was placed in a proneposition. The dorsum was shaved and cleansed. A series of partialthickness wounds (approximately 4 cm²) was created on the dorsum using adermatome set at 0.015 inches in depth. The wounds were placed on theback in mirror image array so that each treatment site had it ownvehicle control site. The reason for mirror image wound placement was tocontrol for the variability of healing that occurs depending upon theanatomic location of the wound.

Immediately after wounding, pseudopterosin A methyl ether ointment wasapplied as a single application to the wounds in concentrations of0.05%, 0.15% and 0.5%. Vehicle alone, and untreated wounds were alsostudied. Ointment was applied such that it evenly covered the woundarea. Following treatment, all wounds were covered with a semi-permeableocclusive dressing (OpSite) followed by a bolster dressing. Photographsand elliptical biopsies (including normal tissue margins) were taken ondays three and eight post-wounding. An occlusive dressing was reappliedafter day 3. Biopsies were bisected. One part was fixed in formalin,embedded in paraffin for routine histologic staining by hematoxylin andcosin (H&E). The second part was snap frozen and cryosat sections madefor immunohistochemical reaction with the proliferative marker Ki-67.

The percent of re-epithelialization of each wound was quantitated byusing a reticle grid in the eyepiece of a Labophot Nikon microscope. Theproliferative activity was quantitated by immunohistochemical stainingof frozen tissue sections with Ki-67 following by scoring of Ki-67immunoreactive cells.

As expected, wounds treated with the vehicle alone, showed very littlere-epithelialization (1.4±11.6%) on day three post-wounding (The Table).Within three days after a single application of pseudopterosin A methylether to individual wounds however, differences in the percentre-epithelialization across the wound were of served. There was a dosedependent, increase in the percent re-epithelialization of woundstreated with increasing concentrations of pseudopterosin A methyl ethersuggesting an acceleration in the rate of healing of such wounds (TheTable).

By day eight all wounds (including vehicle-treated wounds) were healedand showed similar histologic appearance.

                  TABLE                                                           ______________________________________                                        Percent Re-epithelialization of Partial Thickness                             Wounds Three Days After Treatment with a Single                               Application of Pseudopterosin A Methyl Ether                                  Dose of Pseudopterosin A                                                                      Percent Re-epithelialization                                  Methyl Ether (%)                                                                              (Mean % + SEM)                                                ______________________________________                                        0 (vehicle control)                                                                            14 ± 11.6                                                 0.05            14 ± 7.3                                                   0.15            27 ± 8.0                                                   0.5             48 ± 9,7                                                   ______________________________________                                    

The data presented here suggest that a single treatment withpseudopterosin A methyl ether may accelerate the rate of partialthickness wound healing.

Equivalents

Those skilled in the art will know, or be able to ascertain using nomore than routine experimentation, many equivalents to the specificembodiments of the invention described herein. These and all otherequivalents are intended to be encompassed by the following claims.

What is claimed is:
 1. A method of promoting the re-epithelization of apartial thickness wound on an individual or animal comprising contactingthe wound with a composition comprising an effective wound healingamount of a compound having the following structural formula: ##STR15##wherein: R1 and R2 are independently selected from the group consistingof --H, lower alkyl, substituted lower alkyl, phenyl, substitutedphenyl, benzyl, substituted benzyl, acyl, alkoxycarbonyl and amonosaccharide and wherein R1 and R2 are different and one of R1 and R2is a monosaccharide; andR3 is a substituted or unsubstituted lower alkylgroup.
 2. The method of claim 1 wherein the compound has the followingstructural formula: ##STR16## wherein R1 has the following structuralformula: ##STR17## R2 is selected from the group consisting of --H,lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m) --CO--NH₂,--(CH₂)_(m) -(benzyl) and --(CH₂)_(m) -(substituted benzyl), wherein mis an integer from 1 to about 9;R3 is --CH═C(CH₃)₂ ; R4, R5 and R6 areindependently selected from the group consisting of --H and acetyl; andR7 is selected from the group consisting of --H, --CH₃ and --CH₂ OH. 3.The method of claim 1 wherein the compound has the following structuralformula: ##STR18## wherein R1 is selected from the group consisting of--H, lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m) --CO--NH₂,--(CH₂)_(m) -(benzyl) and --(CH₂)_(m) -(substituted benzyl), wherein mis an integer from 1 to about 9;R2 has the following structural formula:##STR19## R3 is --CH═C(CH₃)₂ ; R4, R5 and R6 are independently selectedfrom the group consisting of --H and acetyl; and R7 is selected from thegroup consisting of --H, --CH₃ and --CH₂ OH.
 4. The method of claim 1wherein the compound is selected from the group consisting ofpseudopterosin A, pseudopterosin A methyl ether, pseudopterosin C,pseudopterosin C methyl ether, pseudopterosin E and pseudopterosin Emethyl ether.
 5. A method for reducing the scarring associated with thehealing of a wound on an individual or animal comprising contacting thewound with a composition comprising an effective wound healing amount ofa compound having the following structural formula: ##STR20## wherein:R1 and R2 are independently selected from the group consisting of --H,lower alkyl, substituted lower alkyl, phenyl, substituted phenyl,benzyl, substituted benzyl, acyl, alkoxycarbonyl and a monosaccharideand wherein R1 and R2 are different and one of R1 and R2 is amonosaccharide; andR3 is a substituted or unsubstituted lower alkylgroup.
 6. The method of claim 5 wherein the compound has the followingstructural formula: ##STR21## wherein R1 has the following structuralformula: ##STR22## R2 is selected from the group consisting of --H,lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m) --CO--NH₂,--(CH₂)_(m) -(benzyl) and --(CH₂)_(m) -(substituted benzyl), wherein mis an integer from 1 to about 9;R3 is --CH═C(CH₃)₂ ; R4, R5 and R6 areindependently selected from the group consisting of --H and acetyl; andR7 is selected from the group consisting of --H, --CH₃ and --CH₂ OH. 7.The method of claim 5 wherein the compound has the following structuralformula: ##STR23## wherein R1 is selected from the group consisting of--H, lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m) --CO--NH₂,--(CH₂)_(m) -(benzyl) and --(CH₂)_(m) -(substituted benzyl), wherein mis an integer from 1 to about 9;R2 has the following structural formula:##STR24## R3 is --CH═C(CH₃)₂ ; R4, R5 and R6 are independently selectedfrom the group consisting of --H and acetyl; and R7 is selected from thegroup consisting of --H, --CH₃ and --CH₂ OH.
 8. The method of claim 5wherein the compound is selected from the group consisting ofpseudopterosin A, pseudopterosin A methyl ether, pseudopterosin C,pseudopterosin C methyl ether, pseudopterosin E and pseudopterosin Emethyl ether.
 9. A method of promoting the healing of an internal tissuewound in an individual or animal comprising contacting the wound with acomposition comprising an effective wound healing amount of a compoundhaving the following structural formula: ##STR25## wherein: R1 and R2are independently selected from the group consisting of --H, loweralkyl, substituted lower alkyl, phenyl, substituted phenyl, benzyl,substituted benzyl, acyl, alkoxycarbonyl and a monosaccharide andwherein R1 and R2 are different and one of R1 and R2 is amonosaccharide; andR3 is a substituted or unsubstituted lower alkylgroup.
 10. The method of claim 9 wherein the compound has the followingstructural formula: ##STR26## wherein R1 has the following structuralformula: ##STR27## R2 is selected from the group consisting of --H,lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m) --CO--NH₂,--(CH₂)_(m) -(benzyl) and --(CH₂)_(m) -(substituted benzyl), wherein mis an integer from 1 to about 9;R3 is --CH═C(CH₃)₂ ; R4, R5 and R6 areindependently selected from the group consisting of --H and acetyl; andR7 is selected from the group consisting of --H, --CH₃ and --CH₂ OH. 11.The method of claim 9 wherein the compound has the following structuralformula: ##STR28## wherein R1 is selected from the group consisting of--H, lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m) --CO--NH₂,--(CH₂)_(m) -(benzyl) and --(CH₂)_(m) -(substituted benzyl), wherein mis an integer from 1 to about 9;R2 has the following structural formula:##STR29## R3 is --CH═C(CH₃)₂ ; R4, R5 and R6 are independently selectedfrom the group consisting of --H and acetyl; and R7 is selected from thegroup consisting of --H, --CH₃ and --CH₂ OH.
 12. The method of claim 9wherein the compound is selected from the group consisting ofpseudopterosin A, pseudopterosin A methyl ether, pseudopterosin C,pseudopterosin C methyl ether, pseudopterosin E and pseudopterosin Emethyl ether.
 13. A method of promoting the healing of a wound to dentaltissue or eye tissue on an individual or animal comprising contactingthe wound with a composition comprising an effective wound healingamount of a compound having the following structural formula: ##STR30##wherein: R1 and R2 are independently selected from the group consistingof --H, lower alkyl, substituted lower alkyl, phenyl, substitutedphenyl, benzyl, substituted benzyl, acyl, alkoxycarbonyl and amonosaccharide and wherein R1 and R2 are different and one of R1 and R2is a monosaccharide; andR3 is a substituted or unsubstituted lower alkylgroup.
 14. A method for increasing the adherence of a skin autograft toa wound bed on an individual or animal, comprising applying to the skinautograft an effective wound healing amount of a compound having thefollowing structure formula: ##STR31## wherein: R1 and R2 areindependently selected from the group consisting of --H, lower alkyl,substituted lower alkyl, phenyl, substituted phenyl, benzyl, substitutedbenzyl, acyl, alkoxycarbonyl and a monosaccharide and wherein R1 and R2are different and one of R1 and R2 is a monosaccharide; andR3 is asubstituted or unsubstituted lower alkyl group.
 15. The method of claim14 wherein the compound has the following structural formula: ##STR32##wherein R1 is selected from the group consisting of --H, lower alkyl,benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m) --CO--NH₂, --(CH₂)_(m)-(benzyl) and --(CH₂)_(m) -(substituted benzyl), wherein m is an integerfrom 1 to about 9;R2 has the following structural formula: ##STR33## R3is --CH═C(CH₃)₂ ; R4, R5 and R6 are independently selected from thegroup consisting of --H and acetyl; and R7 is selected from the groupconsisting of --H, --CH₃ and --CH₂ OH.
 16. A method of promoting thehealing of a lesion on the skin of an individual or animal comprisingcontacting the wound with a composition comprising an effective woundhealing amount of a compound having the following structural formula:##STR34## wherein: R1 and R2 are independently selected from the groupconsisting of --H, lower alkyl, substituted lower alkyl, phenyl,substituted phenyl, benzyl, substituted benzyl, acyl, alkoxycarbonyl anda monosaccharide and wherein R1 and R2 are different and one of R1 andR2 is a monosaccharide; andR3 is a substituted or unsubstituted loweralkyl group.
 17. The method of claim 16 wherein the compound has thefollowing structural formula: ##STR35## wherein R1 has the followingstructural formula: ##STR36## R2 is selected from the group consistingof --H, lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH, --(CH₂)_(m)--CO--NH₂, --(CH₂)_(m) -(benzyl) and --(CH₂)_(m) -(substituted benzyl),wherein m is an integer from 1 to about 9;R3 is --CH═C(CH₃)₂ ; R4, R5and R6 are independently selected from the group consisting of --H andacetyl; and R7 is selected from the group consisting of --H, --CH₃ and--CH₂ OH.
 18. The method of claim 16 wherein the compound has thefollowing structural formula: ##STR37## wherein R1 is selected from thegroup consisting of --H, lower alkyl, benzyl, --(CH₂)_(m) --CH₂ OH,--(CH₂)_(m) --CO--NH₂, --(CH₂)_(m) -(benzyl) and --(CH₂)_(m)-(substituted benzyl), wherein m is an integer from 1 to about 9;R2 hasthe following structural formula: ##STR38## R3 is --CH═C(CH₃)₂ ; R4, R5and R6 are independently selected from the group consisting of --H andacetyl; and R7 is selected from the group consisting of --H, --CH₃ and--CH₂ OH.
 19. The method of claim 16 wherein the compound is selectedfrom the group consisting of pseudopterosin A, pseudopterosin A methylether, pseudopterosin C, pseudopterosin C methyl ether, pseudopterosin Eand pseudopterosin E methyl ether.
 20. A method of promoting the healingof a wound on an individual or animal comprising contacting the woundwith a composition comprising an effective wound healing amount of apseudopterosin or a pseudopterosin derivative.
 21. The method of claim20 wherein the wound is a dermal ulcer, a burn wound or a chemical burnwound, an excisional wound on the skin, an internal tissue wound, awound to dental tissue or to eye tissue or a partial thickness wound.22. The method of claim 21 wherein the excisional wound or burn woundextends partially through the epidermis.
 23. The method of claim 21wherein the excisional wound or burn wound extends through at least partof the dermis.
 24. The method of claim 20 further comprising reducingthe scarring associated with the healing of the wound.
 25. A method ofpromoting the growth and proliferation of keratinocytes, endothelialcells and fibroblasts in a wound on an individual or animal or there-epithelization of a partial thickness wound on an individual oranimal comprising contacting the wound with a composition comprising aneffective wound healing amount of a pseudopterosin or a pseudopterosinderivative.
 26. A method of increasing the adherence of a skin autograftto a wound bed on an individual or animal comprising applying to theskin autograft an effective wound healing amount of a pseudopterosin ora pseudopterosin derivative.